Objective:To investigate effects and underlying mechanisms of glutathione persulfate (GSSH) on the level of testosterone in male obese mice.Methods:Totally 45 mice were divided into 3 groups on average. Low-fat diet (LFD)+normal saline (NS) group: 15 mice were fed with LFD for 10 weeks, followed by LFD together with daily intraperitoneal injection of saline for 45 d; high-fat diet (HFD)+NS group: 15 mice were fed with high-fat diet for 10 weeks, followed by HFD and daily intraperitoneal injection of NS for 45 d; HFD+GSSH group: 15 mice were fed with HFD for 10 weeks, followed by a HFD for 45 d and daily intraperitoneal injection of GSSH (200 mg/kg). After the treatment, all mice were killed with their necks-severed, testis and serum were taken out from the mice. Serum levels of testosterone and malondialdehyde (MDA), the mRNA levels of key enzymes for testosterone synthesis ( StAR, 3β- HSD, Cyp11a1 and Cyp17a1) were measured by RT-PCR. The testicular protein levels of StAR, 3β-HSD, NR5A1 and EHD3 were measured by Western blotting assay. Protein levels of NR5A1, SOD and Nrf2 were measured in mouse Leydig TM-3 cells that were treated with 50 μmol/L and 100 μmol/L GSSH, respectively, following with treatment with 100 μmol/L H 2O 2 . Results:1) After treatment, the body weight of mice in HFD+GSSH group did not change significantly, while the body weight of mice in HFD+NS group raised by 24.53% (from 32.46 g to 40.43 g) during the 45-day-intraperitoneal injection ( P=0.002). 2) Serum level of testosterone in HFD+NS group [(12.9±1.7) μg/L] was significantly lower than that in LFD+NS group [(18.3±1.2) μg/L, P=0.020]. However, serum level of testosterone in HFD+GSSH group was (25.42±2.1) μg/L, which was significantly higher than that in HFD+NS group ( P=0.030). The RT-PCR test results showed that compared with LFD+NS group, the expression levels of all key genes involved in testosterone synthesis ( StAR, 3β- HSD, Cyp11a1, Cyp17a1) showed a significant decrease in HFD+NS group ( P=0.003, P=0.007, P<0.001, P<0.001). The expression levels of these genes were restored in the mouse testes of HFD+GSSH group ( P=0.002, P<0.001, P<0.001, P=0.006). 3) Similarly, compared with LFD+NS group [(9.00±1.59) nmol/mL], the serum MDA level of HFD+NS group [(10.61±1.73) nmol/mL] raised significantly ( P=0.016), while GSSH reversed the raised HFD+NS high level of serum MDA in HFD+GSSH group [(9.23±0.94) nmol/mL, P=0.048]. 4) Both levels of NR5A1, EHD3, StAR, and 3β-HSD were reduced in HFD+NS group ( P=0.002, P=0.012, P=0.004, P=0.043), but their levels were significantly restored in HFD+GSSH group ( P<0.001, P=0.017, P=0.004, P<0.001). 5) The levels of NR5A1, Nrf2 and SOD were obviously down-regulated in TM3 cells treated with H 2O 2 ( P<0.001, P=0.002, P=0.004). Conclusion:GSSH can raise serum level of testosterone in HFD-fed mice by up-regulating expression of genes which are important for testicular testosterone biosynthesis.

Objective:To investigate effects and underlying mechanisms of glutathione persulfate (GSSH) on the level of testosterone in male obese mice.Methods:Totally 45 mice were divided into 3 groups on average. Low-fat diet (LFD)+normal saline (NS) group: 15 mice were fed with LFD for 10 weeks, followed by LFD together with daily intraperitoneal injection of saline for 45 d; high-fat diet (HFD)+NS group: 15 mice were fed with high-fat diet for 10 weeks, followed by HFD and daily intraperitoneal injection of NS for 45 d; HFD+GSSH group: 15 mice were fed with HFD for 10 weeks, followed by a HFD for 45 d and daily intraperitoneal injection of GSSH (200 mg/kg). After the treatment, all mice were killed with their necks-severed, testis and serum were taken out from the mice. Serum levels of testosterone and malondialdehyde (MDA), the mRNA levels of key enzymes for testosterone synthesis ( StAR, 3β- HSD, Cyp11a1 and Cyp17a1) were measured by RT-PCR. The testicular protein levels of StAR, 3β-HSD, NR5A1 and EHD3 were measured by Western blotting assay. Protein levels of NR5A1, SOD and Nrf2 were measured in mouse Leydig TM-3 cells that were treated with 50 μmol/L and 100 μmol/L GSSH, respectively, following with treatment with 100 μmol/L H 2O 2 . Results:1) After treatment, the body weight of mice in HFD+GSSH group did not change significantly, while the body weight of mice in HFD+NS group raised by 24.53% (from 32.46 g to 40.43 g) during the 45-day-intraperitoneal injection ( P=0.002). 2) Serum level of testosterone in HFD+NS group [(12.9±1.7) μg/L] was significantly lower than that in LFD+NS group [(18.3±1.2) μg/L, P=0.020]. However, serum level of testosterone in HFD+GSSH group was (25.42±2.1) μg/L, which was significantly higher than that in HFD+NS group ( P=0.030). The RT-PCR test results showed that compared with LFD+NS group, the expression levels of all key genes involved in testosterone synthesis ( StAR, 3β- HSD, Cyp11a1, Cyp17a1) showed a significant decrease in HFD+NS group ( P=0.003, P=0.007, P<0.001, P<0.001). The expression levels of these genes were restored in the mouse testes of HFD+GSSH group ( P=0.002, P<0.001, P<0.001, P=0.006). 3) Similarly, compared with LFD+NS group [(9.00±1.59) nmol/mL], the serum MDA level of HFD+NS group [(10.61±1.73) nmol/mL] raised significantly ( P=0.016), while GSSH reversed the raised HFD+NS high level of serum MDA in HFD+GSSH group [(9.23±0.94) nmol/mL, P=0.048]. 4) Both levels of NR5A1, EHD3, StAR, and 3β-HSD were reduced in HFD+NS group ( P=0.002, P=0.012, P=0.004, P=0.043), but their levels were significantly restored in HFD+GSSH group ( P<0.001, P=0.017, P=0.004, P<0.001). 5) The levels of NR5A1, Nrf2 and SOD were obviously down-regulated in TM3 cells treated with H 2O 2 ( P<0.001, P=0.002, P=0.004). Conclusion:GSSH can raise serum level of testosterone in HFD-fed mice by up-regulating expression of genes which are important for testicular testosterone biosynthesis.

Objective To evaluate the role of circular RNA protein arginine methyltransferase 5 (circPRMT5) in the genesis and progression of colorectal cancer.Methods From January 2013 to December 2017,96 patients with colorectal cancer who underwent radical resection in Department of General Surgery,Jiading District Central Hospital Affiliated to Shanghai Medical College of Health were collected.The expression of circPRMT5 in colorectal cancer tissues was examined by real-time polymerase chain reaction (RT-PCR).The correlation between circPRMT5 expression level and age,gender,tumor size,tumor location,pathological differentiation,TNM stage,lymph node metastasis of patients with colorectal cancer was analyzed.The SW620 and LOVO cells were divided into control group,circPRMT5-lenti group and circPRMT5-shRNA-lenti group according to different interventions.The effects of circPRMT5 expression level on viability,apoptosis,mitochondrial membrane potential and migration of SW620 and LOVO cells were detected.The influence of circPRMT5 expression level on E-cadherin,Slug,N-cadherin and vimentin was determined by Western blotting method.The potential target miRNA of circPRMT5 was predicted by Starbase V2.0.Student's t test,analysis of variance and chi-square test were performed for statistical analysis.Results The results of RT-PCR showed that the expression of circPRMT5 in colorectal cancer tissues was higher than that of adjacent cancer tissues (2.167 ± 0.345 vs.1.103 ± 0.144),and the difference was statistically significant (t =26.847,P ＜ 0.01).The circPRMT5 expression level was positively correlated with tumor size,TNM stage,lymph node metastasis and distant metastasis (x2 =6.010,10.971,5.321 and 6.272,all P ＜0.05).The upregulation of circPRMT5 expression could promote proliferation and migration of SW620 and LOVO cells.The circPRMT5 downregulation could inhibit cell proliferation,induce apoptosis and decrease mitochondrial membrane potential.The results of Western blotting indicated that,compared with those of control group,the expression of Slug,N-cadherin and vimentin increased in circPRMT5-1enti group (1.023 ±0.038 vs.2.105 ±0.042,1.051 ±0.309 vs.2.277 ± 0.111,1.055 ± 0.040 vs.2.002 ± 0.537,respectively),however the expression of E-cadherin decreased (2.074 ± 0.214 vs.0.627 ± 0.023),and the differences were statistically significant (t =31.817,22.065,14.536 and 9.148,all P ＜ 0.01).Compared with the control group,the expression of Slug,N-cadherin and vimentin decreased in circPRMT5-shRNA-lenti group (1.023 ± 0.038 vs.0.585 ± 0.023,1.051 ± 0.309 vs.0.616 ± 0.043,1.055 ±0.040 vs.0.537 ±0.022),while the expression of E-cadherin increased (2.074 ± 0.214 vs.2.756 ± 0.148),and the differences were statistically significant (t =-13.795,-14.252,-11.794 and-13.116,all P ＜ 0.05).A total of 21 miRNAs might have potential binding sites with circPRMT5 predicted by Starbase V2.0 software.The expression of miRNA4735-3p,miRNA202-3p,miRNA326,let-7i-5p and miRNA4500 was negatively correlated with circPRMT5 expression in both SW620 and LOVO cells confirmed by RT-PCR.Conclusion CircPRM75 is an important oneogenic gene in the genesis and progress of colorectal cancer,and may have certain potential application prospect in the research and development for colorectal cancer.

Objective To observe the characteristics of collateral circulation blood flow of ipsilateral ophthalmic artery in patients with internal carotid artery occlusion. Methods The imaging data of 20 patients with internal carotid artery occlusion were analyzed retrospectively. There were 11 males and 9 females, aged from 30 to 65 years, with an average age of (45±3) years. All the patients underwent digital subtraction angiography and transcranial Doppler examination, and 6 patients underwent simultaneous magnetic resonance angiography. The blood supply and collateral circulation of the ipsilateral ophthalmic artery were observed . Results All the patients had unilateral internal carotid artery occlusion. The blood supply of the ipsilateral internal carotid artery and ophthalmic artery comes from the collateral circulation between the middle meningeal artery branches of the external carotid artery and the ophthalmic artery in 18 patients (90.0%); it also comes from the anterior communicating artery of the contralateral internal carotid artery in 16 patients (80.0%); and the posterior communicating artery of the contralateral internal carotid artery in 12 patients (60.0%), respectively. Conclusion The blood flow of the ipsilateral ophthalmic artery mainly comes from the middle meningeal artery branch of the ipsilateral external carotid artery, also comes from the anterior and posterior communicating arteries of the contralateral internal carotid artery.

Objective To investigate the clinical effect of Ethibond suture and anchor fixation by arthroscopic technique for the treatment of avulsion fracture of the anterior cruciate ligament (ACL) tibial insertion.Methods Twenty patients with avulsion fracture of the ACL tibial insertion hospitalized between July 2013 and June 2014 were collected retrospectively.There were 12 males and 8 females,aged 18-41 years (mean,25.3 years).All patients were identified with type Ⅱ and type Ⅲ fractures according to the Meyers-McKeever classification.Results of Lachman test and anterior drawer test were both positive.All patients accepted the operation that avulsion fracture of the ACL tibial insertion was treated with the Ethibond suture and anchor fixation by arthroscopic technique within 3 weeks after injury.Follow-up X-ray examinations were carried out to evaluate the bone union.Lysholm and international knee documentation committee (IKDC) scores were used to evaluate the function of knee joint postoperatively.Results Operation time was 45-70 min (mean,50 min).Blood loss was 5-15 ml (mean,10 ml).Follow-up was conducted for 12-24 months (mean,16.3 months).Postoperative X-ray showed the reduction was satisfactory.Lachman test and anterior drawer test were both negative after operation.Knee functions were recovered to normal.Lysholm and IKDC scores were 89-96 points [(93.5 ± 2.3) points]and 84-96 points [(91.0 ± 3.9)points] respectively at the final follow-up,improved compared to the preoperative 42-61 points [(51.1 ± 6.2) points] and 46-68 points [(55.2 ± 7.0) points] (both P ＜0.05).Conclusion Arthroscopic Ethibond suture and anchor fixation for avulsion fracture of the ACL tibial insertion is an effective procedure with the advantages of small trauma,reliable fixation,good functional recovery and satisfactory clinical effect.

Objective To investigate the effects of IL-1 β on proliferation and migration of gallbladder cancer cells.Methods The secretion of IL-1 β in tissues of gallbladder cancer, chronic cholecystitis and normal gallbladder as well as in supernatant of gallbladder cancer cell lines (GBC-SD, SGC996) and HIBEpic cells was determined by enzyme-linked immunosorbent assay (ELISA) method.The levels of IL-1 β mRNA in GBC-SD, SGC996 and HIBEpic cells were measured by RT-PCR assay.The effects of exogenous IL-1 β on the proliferation of GBC-SD and SGC996 cells in vitro and in vivo were evaluated using WST-1 assay and xenograft tumor model, respectively.The effects of exogenous IL-1 β on the migration of GBC-SD and SGC996 cells in vitro were measured by Tranwell assay.The levels of Twist protein in GBC-SD and SGC996 ceils were examined by western blot assay after treatment with exogenous IL-1 β.In addition, the proliferation and migration of GBC-SD and SGC996 cells after gene silencing of Twist by Twist-siRNA were also evaluated.Results The level of IL-1β protein in normal gallbladder was low (66.4 ± 35.0)pg/ml,while it was significantly increased in gallbladder cancer and chronic cholecystitis [(616.4 ± 95.7) pg/ml and (422.3 ± 48.9) pg/ml, P ＜ 0.05].The levels of IL-1 βin GBC-SD and SGC996 cell culture medium [(587.4 ± 99.8) pg/ml and (657.2 ± 76.6) pg/ml] were much higher than those in the HIBEpic cells [(38.4± 12.1)pg/ml, P ＜ 0.05].Exogenous IL-1β promoted the proliferation of GBC-SD and SGC996 cells both in vitro and in vivo as well as migration in vitro (P ＜ 0.05).The level of Twist protein was significantly increased after treatment with exogenous IL-1 β.In addition, gene silencing of Twist blocked IL-1 β-induced proliferation and migration of GBC-SD and SGC996 cells.Conclusion IL-1 β promoted proliferation and migration of gallbladder cancer cells via Twist activation.